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PMID: 15004232 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification and characterization of two novel proteins affecting fission yeast gamma-tubulin complex function.

Molecular biology of the cell ·Vol. 15 ·No. 5 ·2004-05-00 ·Pages 2287-301

Venkatram S, Tasto JJ, Feoktistova A, Jennings JL, Link AJ, Gould KL

Abstract

The gamma-tubulin complex, via its ability to organize microtubules, is critical for accurate chromosome segregation and cytokinesis in the fission yeast, Schizosaccharomyces pombe. To better understand its roles, we have purified the S. pombe gamma-tubulin complex. Mass spectrometric analyses of the purified complex revealed known components and identified two novel proteins (i.e., Mbo1p and Gfh1p) with homology to gamma-tubulin-associated proteins from other organisms. We show that both Mbo1p and Gfh1p localize to microtubule organizing centers. Although cells deleted for either mbo1(+) or gfh1(+) are viable, they exhibit a number of defects associated with altered microtubule function such as defects in cell polarity, nuclear positioning, spindle orientation, and cleavage site specification. In addition, mbo1Delta and gfh1Delta cells exhibit defects in astral microtubule formation and anchoring, suggesting that these proteins have specific roles in astral microtubule function. This study expands the known roles of gamma-tubulin complex components in organizing different types of microtubule structures in S. pombe.

MeSH Terms
Binding Sites Calmodulin-Binding Proteins/analysis,genetics,metabolism Cell Division/genetics,physiology Cell Polarity/genetics Gene Deletion Green Fluorescent Proteins Intranuclear Space/ultrastructure Microtubule-Associated Proteins/genetics,metabolism,physiology Microtubule-Organizing Center/metabolism,physiology Microtubules/metabolism,physiology Models, Biological Molecular Sequence Data Protein Interaction Mapping Schizosaccharomyces/genetics,metabolism,ultrastructure Schizosaccharomyces pombe Proteins/genetics,metabolism,physiology Sequence Alignment Spindle Apparatus/genetics,metabolism Tubulin/metabolism
Chemicals
Calmodulin-Binding Proteins Gfh1 protein, S pombe Mbo1 protein, S pombe Microtubule-Associated Proteins Schizosaccharomyces pombe Proteins Tubulin enhanced green fluorescent protein Green Fluorescent Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Venkatram Srinivas
Department of Cell and Developmental Biology, Howard Hughes Medical Institute, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Tasto Joseph J
Feoktistova Anna
Jennings Jennifer L
Link Andrew J
Gould Kathleen L
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-10-0728
Published
2004-05-00
Epub
2004-00-05
Pages
2287-301
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC404023
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.e03-10-0728 PMC Full Text View journal details
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